The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization.
The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization.
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DOI：
10.3390/jof6030151
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发表时间：
2020-08-26
期刊：
影响因子：
--
通讯作者：
Aimanianda V
中科院分区：
文献类型：
--
作者：
Valsecchi I;Stephen-Victor E;Wong SSW;Karnam A;Sunde M;Guijarro JI;Rodríguez de Francisco B;Krüger T;Kniemeyer O;Brown GD;Willment JA;Latgé JP;Brakhage AA;Bayry J;Aimanianda V
烟曲霉分生孢子的免疫惰性归因于其表面由RodAp组成的棒状层，其特征是8个保守的半胱氨酸残基形成4个二硫键。早些时候，我们发现保守的半胱氨酸残基点(CCRP)突变导致分生孢子没有小棒层。在这里，我们扩展了我们的研究，比较了携带ccrp突变的分生孢子和缺失突变体(∆roda)的分生孢子的表面组织和免疫反应性。用抗RodAp抗体进行的Western印迹分析表明，CCRP突变的分生孢子的细胞质中没有RodAp。免疫标记显示对分生孢子表面葡聚糖有不同的反应性，ccrp突变体分生孢子优先结合α-(1，3)-葡聚糖，∆RodA分生孢子选择性结合β-(1，3)-葡聚糖，亲本菌株分生孢子呈阴性标记。然而，ccrp突变体和∆roda的透性与亲本分生孢子相似。Ccrp突变体的分生孢子表面暴露蛋白的蛋白质组学分析表明，它们与亲本菌株有更多的相似性，但与∆RodA有显著差异。与∆RodA分生孢子相比，Ccrp突变分生孢子的免疫刺激作用较弱。我们的数据表明：(I)保守的半胱氨酸残基对RodAp的运输和分生孢子表面小棒层的组织是必不可少的，(Ii)定点突变可能是研究真菌细胞壁基因在宿主-真菌相互作用中作用的另一种方法。
Immune inertness of Aspergillus fumigatus conidia is attributed to its surface rodlet-layer made up of RodAp, characterized by eight conserved cysteine residues forming four disulfide bonds. Earlier, we showed that the conserved cysteine residue point (ccrp) mutations result in conidia devoid of the rodlet layer. Here, we extended our study comparing the surface organization and immunoreactivity of conidia carrying ccrp-mutations with the RODA deletion mutant (∆rodA). Western blot analysis using anti-RodAp antibodies indicated the absence of RodAp in the cytoplasm of ccrp-mutant conidia. Immunolabeling revealed differential reactivity to conidial surface glucans, the ccrp-mutant conidia preferentially binding to α-(1,3)-glucan, ∆rodA conidia selectively bound to β-(1,3)-glucan; the parental strain conidia showed negative labeling. However, permeability of ccrp-mutants and ∆rodA was similar to the parental strain conidia. Proteomic analyses of the conidial surface exposed proteins of the ccrp-mutants showed more similarities with the parental strain, but were significantly different from the ∆rodA. Ccrp-mutant conidia were less immunostimulatory compared to ∆rodA conidia. Our data suggest that (i) the conserved cysteine residues are essential for the trafficking of RodAp and the organization of the rodlet layer on the conidial surface, and (ii) targeted point mutation could be an alternative approach to study the role of fungal cell-wall genes in host–fungal interaction.
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影响因子：
3.6
作者：
Gilbert NM;Donlin MJ;Gerik KJ;Specht CA;Djordjevic JT;Wilson CF;Sorrell TC;Lodge JK
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